中华麻醉学杂志
中華痳醉學雜誌
중화마취학잡지
CHINESE JOURNAL OF ANESTHESIOLOGY
2014年
12期
1495-1497
,共3页
异氟醚%缺血后处理%缺氧%氧%大脑皮质%神经元%凋亡调节蛋白质类
異氟醚%缺血後處理%缺氧%氧%大腦皮質%神經元%凋亡調節蛋白質類
이불미%결혈후처리%결양%양%대뇌피질%신경원%조망조절단백질류
Isoflurane%Ischemic postconditioning%Anoxia%Oxygen%Cerebral cortex%Neurons%Apoptosis regulatory proteins
目的 评价异氟醚后处理对大鼠缺氧缺糖-复氧复糖皮质神经元促凋亡蛋白表达的影响.方法 体外培养出生24 h内的SD大鼠原代皮质神经元,以1×106个/ml的密度接种于6孔板(2ml/孔)培养皿,采用随机数字表法分为3组(n=12):对照组(C组)神经元正常培养;缺氧缺糖-复氧复糖组(OGD/R组)神经元缺氧缺糖30 min后复糖复氧;异氟醚后处理组(Ⅰ组)神经元缺氧缺糖30 min后复糖复氧即刻向培养皿中通入2％异氟醚,培养1h.于培养24h时收集神经元,采用Hoechst/PI双染色法测定神经元凋亡率;采用免疫印迹法测定cmpme-3蛋白的表达;采用PCR法测定Bid、B im和Puma的mRNA表达;采用Westem blot法测定Bid 、Bim和Puma的蛋白表达.结果 与S组比较,OGD/R组神经元凋亡率升高,caspase-3蛋白、Bid、Bim和Puma的mRNA和蛋白表达上调(P＜0.05);与OGD/R组比较,Ⅰ组神经元凋亡率下降,caspase-3蛋白、Bid、Bim和Puma的mRNA和蛋白表达下调(P＜0.05).结论 异氟醚后处理通过下调促凋亡蛋白表达,抑制大鼠缺氧缺糖-复氧复糖皮质神经元凋亡.
目的 評價異氟醚後處理對大鼠缺氧缺糖-複氧複糖皮質神經元促凋亡蛋白錶達的影響.方法 體外培養齣生24 h內的SD大鼠原代皮質神經元,以1×106箇/ml的密度接種于6孔闆(2ml/孔)培養皿,採用隨機數字錶法分為3組(n=12):對照組(C組)神經元正常培養;缺氧缺糖-複氧複糖組(OGD/R組)神經元缺氧缺糖30 min後複糖複氧;異氟醚後處理組(Ⅰ組)神經元缺氧缺糖30 min後複糖複氧即刻嚮培養皿中通入2％異氟醚,培養1h.于培養24h時收集神經元,採用Hoechst/PI雙染色法測定神經元凋亡率;採用免疫印跡法測定cmpme-3蛋白的錶達;採用PCR法測定Bid、B im和Puma的mRNA錶達;採用Westem blot法測定Bid 、Bim和Puma的蛋白錶達.結果 與S組比較,OGD/R組神經元凋亡率升高,caspase-3蛋白、Bid、Bim和Puma的mRNA和蛋白錶達上調(P＜0.05);與OGD/R組比較,Ⅰ組神經元凋亡率下降,caspase-3蛋白、Bid、Bim和Puma的mRNA和蛋白錶達下調(P＜0.05).結論 異氟醚後處理通過下調促凋亡蛋白錶達,抑製大鼠缺氧缺糖-複氧複糖皮質神經元凋亡.
목적 평개이불미후처리대대서결양결당-복양복당피질신경원촉조망단백표체적영향.방법 체외배양출생24 h내적SD대서원대피질신경원,이1×106개/ml적밀도접충우6공판(2ml/공)배양명,채용수궤수자표법분위3조(n=12):대조조(C조)신경원정상배양;결양결당-복양복당조(OGD/R조)신경원결양결당30 min후복당복양;이불미후처리조(Ⅰ조)신경원결양결당30 min후복당복양즉각향배양명중통입2％이불미,배양1h.우배양24h시수집신경원,채용Hoechst/PI쌍염색법측정신경원조망솔;채용면역인적법측정cmpme-3단백적표체;채용PCR법측정Bid、B im화Puma적mRNA표체;채용Westem blot법측정Bid 、Bim화Puma적단백표체.결과 여S조비교,OGD/R조신경원조망솔승고,caspase-3단백、Bid、Bim화Puma적mRNA화단백표체상조(P＜0.05);여OGD/R조비교,Ⅰ조신경원조망솔하강,caspase-3단백、Bid、Bim화Puma적mRNA화단백표체하조(P＜0.05).결론 이불미후처리통과하조촉조망단백표체,억제대서결양결당-복양복당피질신경원조망.
Objective To evaluate the effect of isoflurane post-conditioningon the expression of pro apoptotic proteins in the conical neurons exposed to oxygen-glucose deprivation and restoration (OGD/R) in rats.Methods Primary cortical neurons isolated from male Sprague-Dawley rats (within 24h after birth),were cultured in vitro and inoculated in 6-well culture plate (2 ml/well) at a density of 1 × 106/ml.The cells were divided into 3 groups (n =12 each) using a random number table:control group(group C),OGD/R group,and isoflurane post-conditioning group (Ⅰ group).In OGD/R group,the cells were incubated in glucose-free BBS aerated with 95 ％ N2 for 30 min followed by restoration of 2-glucose supply for 1 h.At 24 h of incubation,the cells were collected for detection of neuronal apoptosis (.using Hoechst/PI staining),caspase-3 expression (by Western blot),expression of Bid,Bim and Puma mRNA (by PCR),and expression of Bid,Bim and Puma (by Western blot).Apoptosis rate was calculated.Results Compared with S group,the apoptosis rate was significantly increased,and the expression of caspase-3 and Bid,Bim and Puma mRNA and protein was upregulated in OGD/R group.The apoptosis rate and expression of caspase-3 and Bid,Bim and Puma mRNA and protein were significantly lower in Ⅰ group than in OGD/R group.Conclusion Isofluranepost-conditioning inhibits apoptosis in the cortical neurons exposed to OGD/R through down-regulating pm-apoptotic proteins in rats.
